Diastatic substance and method of making same.



ow an JOKICHI TAKAMINE, OF NEW YORK, N. Y.

DIASTATIC SUBSTANCE AND METHOD OF MAKING SAME- Specification of Letters Patent.

Patented July 24, 1906.

Application filed January 21. 1905. Serial No. 242,116.

To all whom it may concern.-

Be it known that I, JOKICHI TAKAMINE, a subject of the Em eror of Japan, residing in the borough of anhattan, in the city,

5 county, and State of New York, have invented and discovered new and useful Diastatic Substances and Method of Producing the Same, of which the following is a specification. a

My invention relates to diastatic substances or enzyms having the property of converting starch or the starchy constituents of grains, roots, tubers, or other portions of vegetable anatomy into sugars.

The object of my invention is to isolate and render useful and available for use in the arts a certain diastatic principle or enzym which I have discovered in the seeds. grains, roots. tubers. o other portions of vegetable structure or composition, and, further, to produce said principle or enzym in a ermanent stable form,- in which it may be l ept or stored without deteriorating or losing its properties. 1

A further object of my invention is to pro- I duce a new diastatic substance having liquefilling and saccharifying properties in which t ese starch iquefied by the liquef'ying properties may be saccharified by the saccharifying principle or so adjusted that the sacchari 1ng properties of the diastase are not in excess.

It is well known that diastatic enzyms in general possess the characteristic roperty of converting starch into sugars. his conversion passes through various distinctive sta' es.

irst. The first is the li uefying stage, wherein the starch is liquefie or dissolved:

Second. The next stage is the dextri stage, in which the liquefied or disso ved starch is transformed into dextrin. This is a transition stage and for the pu oses of this specification may be left out of rther consideration.

Third. The final stage is the saccharifyin stage, in which .the dextrin is transformed into sugars.

I have discovered that these se arate and distinct stages in the conversion 0? starch, or at least the liguefying and saccharif'ying stages, are pro uced by separate and distinct enzymotic agents whic exist together 5 5 in varying proportionsin diastases'heretofore c used." Whereas these different roperties are ad usted so that the enzymotic agents may all be present in known diastases, they do not occur therein in the most advantageous or desirable ropor' tions for the economic conversion 0 starch or starchy materials into sugar. For example; in a certain diastase the proportion of quefying enzym present may be in excess of that required to liquefy the amount of starch which the saccharifying enzym present can convert to sugar. In another diastase the relative proportions of the liquef'ying and saccharifying enzyms may be the reverse of the foregoing example. By my invention the proportions of the liquefying and saccharifying enzyms may be adjusted and a new diastatic substance produced having liquefying and saccharifying enzyms in the most advanta eous proportions for securing the greatest e ciency in result and savingin the material used and wherein the proportion of each enzym is such as to convert completely a given mass of starch or starchy material without there being a surplus of one enzym over the other or any portion of. the

starch remaining unconverted.

' It is well known that diastasesat-pre'sent known and used are produced by the germination of seeds or grains, usually of cereals, the growthbeing conducted under" roper conditions of temperature and humidity. 01

by the growth of certain fungi upon suitable culture media. Diastases so dproduced possess-both starch liquefying an saccharifying properties, and hence cause the transformation of starch into sugars; but such diastases are irregular and variable in the rela-. tive proportions of liquefying and saccharifil-ing enzyms present therein, and .hence t eir action is more or less uncertain, conjectural, and incomplete.

' I have discovered that an enzym having marked saccharif ing diastatic properties may be produced Tom the seeds or grains of cereals which have not been subjected to germination nor to the growth of fungi thereon or from the epidermis or cuticular portions of said grains remaining after the removal of the whole or larger part of the starchy constituents and not subjected to fun 'oid growth. In order to obtain this enzym, f take a suitable quantity of broken or coarsely-ground e. g.,'of bran, shorts, middlings. or the like-- ency its saccharifying properties may be prefound that the saccharifying enzym existing active ingredients SSP and macerate, percolate. or'otherwise extract the same with water at the ordinary temperature-or slightly above. For the sake of coneiseness and to save repetition I employ in the claims the terms grains ant roots as generic expressions to designate materials suitable for use in carrying out the invention, it being understood that such materials are best treated in a broken, ground, or comminuted condition to obtain the fluid extract therefrom. The extract so obtained is a brownish or yellowish liquid, the color being de endent uponthe materials selected.

his extract or the raw material from tract herein described or of the saccharifyin principle isolated or separated therefrom am enabled to produce a general diastase having greatly-increased efficiency in causing the conversion of starch into sugar. This I accomplish by adding or admixing with a diastase or other substance )ossessing starchliquefying properties an deficient in saccharifying properties a proportion of the saccharifying extractor of the isolated principle herein described and obtaining thereb' a diastatic a ent which will convert stare into sugar wit much greater ra idity, completeness, and efliciency than t e ordinary diawhich it is obtained is without effect or with out appreciable effect upon gelatimzed l starch, so that in the ordinary sense ofthe word such extract has no diastatic power, except possibly in very minute quantity of no practical use in the arts. I have discovered,

'owever, that it contains apowerful saccharifving enzym, which may be rendered. available by first liquefying starch with a liquefying diastatic agent or other substance possessing the property of making starch soluble such, for instance, as weak acids. As soon as the starch is in process of liquefaction the extract obtained, as above described, exerts a powerful and rapid saccharifying effect upon it, quickly and effectively causing its transformation into sugars. Thus by-mixing a portion of the extract with a diastatic agent possessing liquefying properties with. respect to starch the conversion of'the starch mto sugar roceeds with greatly-increased rapidity. his can readily be observed by com-- parative tests with the Well-known iodin reaction.

I have found that the extract containing the saccharifying enzym above mentioned deteriorates or loses its power ,on keeping when in the form of a weak solution; but I have discovered that if the said extract be evaporated under reduced pressure or in vacuum until it assumes a syrupy consistserved for a considerable period. I have also in the extract is diflicultly soluble in alcohol, and may, therefore, be separated from'nonof the extract by the addition of a suflicient amount of alcohol. The saccharifying enzym thus obtained when freed from alcohol and dried is a whitish owder and has a remarkable power of sac- :harifying starch which has been reviously liquefied either by the agency of a iquefying enzym or weak acids.

The production of this saccharifying enzym and its isolation and separation constitute one feature of my invention.

Another feature of my invention consists in the application of the said saccharifying enzym to the production of a new and valup eserving and preventingstases used. p

This new diastase may be produced in various ways: To a solution containing an ordinary diastase or a substance possessing starch-liquefying properties, but deficient in saccharifying properties, is added a portion of the extract herein described sufiieient to saccharify all the starch which the liquefying properties of the first diastase will liq'uefy or render soluble. This proportion is dependent upon the liquefying capacity of the selected diastase or substance having liquefying pro erties" and is readily determined by the skil ed worker. The above solution thus made may be evaporated to'a syrupy condition or to a dry mass under reduced pressure, or a dry diastatic agent of improved character in which the proportions of liquefying and saccharifying principles are properly balanced, as stated above, may be obtained by precipitating the mixed fluid with alcohol and separating and drying the precipitate. This precipitate constitutes a diastatic agent with both liquefying and saccharifying principles and of much more vigorous, complete, and efficient action than the original diastases and will much more rapidly and thoroughly convert a given amount of starch to sugar,or suitable proportions of ordinary diastase in dry form may cipitated saccharifying' enzym herem described and a greatly-improved diastase obtained of the same eflicient character, as by the methods above described. 1

i The new diastatic a ent, whether in the form of a fluid or in dry form, and the process of producin the same .and having its saccharifying' iastatic properties composed of or augmented by the saccharifylng diastatic principle'herein described also forms a feature of my invention.

A further feature of my invention consists in the process which I have discovered for c the new sacchari- Tymg diastatic agent herein described from deterioration, whereby the same may be made .Stable and kept or stored indefinitely without losing its properties. I have found that the new diastatic substance above referred to, consisting of a-diastase having saccharifying able diastatic agent. By means of the ex- .diastatic properties imparted to it bythe be mixed with the isolated or prev ceases new saccharifying enzym herein described,

7 orprecipitated principle when departed from this ingredient may be kept and stored without fear of losing its virtues. Various modes to. I have discovered that said ingredient is rendered insoluble and may be precipitated from the extract if an acid condition be induced in the extract or solution. One method of bringing about this acid condition is by fermentation. In order to accomplish this, the extract is subjected to proper fermentin conditions by allowing the same to stand or a period of from thirty to fifty hours, preferably about forty-eight hours, in a warm room at summer temperature-via, about 80 to 90 Fahrenheit. The fermentation sets in with the formation of a whitish precipitate. The precipitate having formed, t e supernatant liquid is drawn off. This liquid contains the saccharifying enzym in a more stable and permanent form than before. It may be concentrated by evaporation at low temperature to a syrupy consistency and preserved. This liquid may then be added in any suitable or desired proportions to a socharifying lution of ordinary diastase or a diastase or other substance possessing starch-liquefying roperties and the mixed liquids precipitated y means of alcohol. The precipitate so obtained possesses both the liquefying and saccharifying diastatic properties in a more stable and permanent form, in which it may be kept or stored without deterioration or loss of its saccharifying powers; or instead of subjecting the extract to the process of fermentation, as above referred to, a similar condition may be brou ht about by the addition of some dilute task .as, for instance, lactic acid. A whitish precipitate similar to that above referred to will be formed and may be removed by filtration or otherwise separated from the liquid. The liquid may then be treated with alcohol to precipitate the sacprinciple therefrom or mixed with a solution fying properties and the Inixe fluid precipitated with alcohol, with the result that a simi lar diastatie substance is produced having both liquefying and saccharifying properties in stable and permanent form not subject to deterioration.

By the above invention I am enabled to produce, and isolate a new and' useful enzym or diastatic agent possessing remarkable sacof separating thisingredient may be resorted "tating from said extract of diastase possessing starch-lique,

charifying properties in a concentrated form, which may be used with diastases heretofore known to greatly increase their efficiency and usefulness and also to produce a diastase possessing the desired proportions of liquefying and saccharifying enzyms and by means of which the amount of diastases at present known which is necessary to accomplish the conversion of a given amount of starchy material into sugar may be greatly reduced,thus

, effect" g a large saving in the cost ofsuch proce and at the same time giving a diastatic agent of definite and known compositi on as regards its liquefying and saccharifying ingredients.

Having thus described my invention and the manner of making and using the same,

what I claim, and desire to protect by Letters Patent, is

1. The process for producing a diastatic substance which consists in making a fluid extract of grains or roots of the character described, or of such materials after the removal therefrom their starchy constituents, precipitating from said extract associated extractive matters which have a deteriorating effect on the stability or permanence thereof, and adding the fluid extract so obtained in desired uantity to a substance haying starch-liqudiying properties.

2. The process for producing a diastatic substance which consists in making a fluid extract of grains, or roots of the character described or of such materials after the removal therefrom of the whole or a portion of their starchy constituents, precipitating from said extract associated extractive matters which have a deteriorating effect on the stability or permanence thereof, treating the fluid extract obtained with alcohol, collecting the precipitate and adding the same in desired quantity to a substance having starch-lique fying properti es.

3. he process of producing a saccharifying diastatic substance having practically no lique ing action on starch, but having marked saccharifying action on liquefied or gelatinized starch, said process consisting in making a fluid extract of grains or roots of the character described, or of such materials after the whole or a portion of their starchy constituents have ben removed and precipiassociated extractive matters having a deteriorating influence on the permanence, stability and keeping qualities of said saccharifying diastatic substance.

4. The process of producing a saccharifying diastatie substance havin practically no liquefying action on stars 1 but having marked saccharifying action on liquefied or gelatinized starch, said process consisting in making a fluidextract of grains or roots of the character described, or of such substances of the whole or a portion of after the whole or a portion of their starchy constituents have been removed, inducing an acid condition in said extract, and se arating the fluid from theprecipitate forme 5. The process of producing a .saccharify ing diastatic substance in a permanent an stable form which consists in making an aqueous extract of grains or roots of the character described, or of such substances after the whole or a portion of their starchy COD-1 stituents have been removed, subjecting said extract to acid fermentation, and removing the precipitate formed.

6. The process of producing a saccharitying diastatic substance in a permanent and stable form which consists in making an aqueous extract of grains or roots of the charjpreeipitate alcohol and i 7. The process of producin acter described, or of such substances after the whole or a portion of their starchy constituents have been removed, subjecting said extract to acid fermentation, removing the formed, treating the fluid with collecting the desired precipitate.

a saccharifyle f0 rm which extract. of

diastatic substance in a sta nsists in making an aqueous -"igi'ains or roots of the character described, or not such substances after the whole or a portion of their starchy constituents have been 'removecl, subjecting said extract to a temperature of from 80 to 90 Fahrenheit for a period of from thirty to; fty hours andremovi'n the precipitate formed.

8. T e herein described. diastatic substance having a whitish color in dry form,

(11 soluble in water, diflicultly soluble in alcohol,

having practically no liquefying action on starch, but having marked sacchar-iiying action on liquefied starch, and substantially free from foreign or associated matters which have a destructive or deteriorating effect on its saccharifying properties.

9. A diastatic substance possessing both starch liquefying and saccharifying properties and in which the saccharifying properties are constituted in Whole or in part by an extrinsic diastatic substance having practically no liquefying action on starch.

10. A diastatic substance in stable form or condition consisting of a mixture of a substance having a liquefying action on gelatinized starch and substant ally no action upon gelatinized starch but having marked saccharifying ac-- tion on liquefied starch or starch solution and freed from deteriorating ingredients.

J OKICHI TAKAMINE.

Witnesses:

HENRY C. WORKMAN, MATHILDA HELEN ANDRES.

a dias'tatic substance having 

